3.1.2.2. Individual rearing approach

  1. Fill standard 1.5 ml Eppendorf® reaction tubes 1/3 full with autoclaved pupation soil (see section 3.1.2.1.). Seal the tubes, place them in laboratory trays, and puncture the lids with a needle (3-4 small holes) to allow air circulation.
  2. Collect wandering larvae from the rearing program (see section 3.1.2.1.).
  3. Gently introduce an individual larva onto the soil of each tube using soft tweezers.
  4. Cover the larvae with another 1/3 full with autoclaved pupation soil (see section 3.1.2.1.).
  5. Seal the tubes.
    -. Follow instructions for storage conditions of trays in incubators until adult emergence (see section 3.1.2.1.).
  6. Transfer adults, which have finished pupation (Fig. 3c), into new individual tubes.
  7. Provide with 1:1 honey water ad libitum using a pipette.
    The SHBs can be maintained for up to 8 weeks in darkness at RT.

Notes:

If any other diet is used to rear or maintain SHB (e.g. to test reproductive success (Ellis et al., 2002b; Buchholz et al., 2008)), the food should be frozen first, especially if field collected (e.g. fruit) to kill any other insect eggs that may be present.